Low Long Noncoding RNA Growth Arrest-Specific Transcript 5 Expression in the Exosomes of Lung Cancer Cells Promotes Tumor Angiogenesis
Figure 6
GAS5 regulated PTEN, PI3K, and AKT by competitively binding miR-29-3p with PTEN. (a) Cytoplasmic and nuclear GAS5 level in vascular HUVECs was detected by qPCR, MT RNR, GAPDH, and U6 served as the internal reference. (b) Cobinding miRNAs for GAS5 and PTEN were predicted by STARBASE 2.0 software. (c) The potential binding sites for GAS5 and PTEN-3′UTR in miR-29a-3p, miR-29b-3p, and miR-29c-3p. (d)–(f) Relative luciferase activity after 48 h cotransfection with the dual luciferase reporter constructs pmiRGlo-GAS5, pmiRGlo-PTEN-3′UTR-1, and pmiRGlo-PTEN-3′UTR-2 (wild-type or mutant vectors) and miR-29a-3p, miR-29b-3p, or miR-29c-3p mimics. NC mimics were used as control group. (g, h) Total PTEN, PI3K, AKT, phosphorylated p-PI3K, and p-AKT levels in the GAS5 overexpression group and GAS5 overexpression combined with cotransfection of miR-29a-3p, miR-29b-3p, or miR-29c-3p were detected by western blot. (i) The expression of miR-29a-3p, miR-29b-3p, and miR-29c-3p in the GAS5 overexpression HUVECs and control groups was detected by qPCR; U6 served as the internal reference. Statistical significance, P<0.05, P<0.01.